Multiplex bacterial growth monitoring in 24-well microplates using a dual optical sensor for dissolved oxygen and pH

Authors

  • Anna S. Kocincová,

    1. University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germany; telephone: +49-941-943-4066; fax: +49-941-943-4064
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  • Stefan Nagl,

    1. University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germany; telephone: +49-941-943-4066; fax: +49-941-943-4064
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  • Sarina Arain,

    1. University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germany; telephone: +49-941-943-4066; fax: +49-941-943-4064
    2. Presens GmbH, Regensburg, Germany
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  • Christian Krause,

    1. Presens GmbH, Regensburg, Germany
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  • Sergey M. Borisov,

    1. University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germany; telephone: +49-941-943-4066; fax: +49-941-943-4064
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  • Matthias Arnold,

    1. DasGip AG, Jülich, Germany
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  • Otto S. Wolfbeis

    Corresponding author
    1. University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germany; telephone: +49-941-943-4066; fax: +49-941-943-4064
    • University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors, D-93040 Regensburg, Germany; telephone: +49-941-943-4066; fax: +49-941-943-4064.
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Abstract

Non-invasive, simultaneous optical monitoring of oxygen and pH during bacterial cultivation in 24-well microplates is presented using an integrated dual sensor for dissolved oxygen and pH values. The dual sensor is based on oxygen-sensitive organosilica microparticles and pH-sensitive microbeads from a polymethacrylate derivative embedded into a polyurethane hydrogel. The readout is based on a phase-domain fluorescence lifetime-based method referred to as modified frequency domain dual lifetime referencing using a commercially available detector system for 24-well microplates. The sensor was used for monitoring the growth of Pseudomonas putida bacterial cultures. The method is suitable for parallelized, miniaturized bioprocessing, and cell-based high-throughput screening applications. Biotechnol. Bioeng. 2008;100: 430–438. © 2008 Wiley Periodicals, Inc.

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